发育中的人胎小肠神经肽的研究

目的　研究人胎小肠壁P物质 (SP)和降钙素基因相关肽 (CGRP) IR肽能神经的分布和发育规律。方法　苏木精伊红 (HE)染色和免疫组织化学ABC法。结果　在胎儿发育过程中 ,小肠组织形态及其神经肽的发生、分化出现明显的变化 ,其分化由十二指肠向空肠、回肠依次进行。第 14周开始 ,胎儿小肠壁粘膜下层、肌层间结缔组织中偶见SP能、CGRP能神经纤维及神经元免疫反应产物 ,第 34周至 38周时反应最强 ,神经元从浅棕色到深棕色 ,神经纤维呈串珠状或点线状。从SP、CGRP免疫组织化学相邻切片上看 ,两者部分存在共存现象。结论　人胎小肠壁存在SP IR、CGRP IR肽能神经 ,粘膜下和肌间神经丛存在SP IR、CGRP IR肽能神经元和神经纤维 ,二种神经肽的分布有差异     

P物质和降钙素基因相关肽在大鼠垂体前叶神经纤维内的共存

为观察P物质(SP)和降钙素基因相关肽(CGRP)在大鼠垂体前叶神经纤维内的共存,用种属特异性抗体(兔抗-SP多克隆抗体和小鼠抗-CGRP单克隆抗体)进行免疫荧光双标记,激光共聚焦扫描显微镜观察进行研究。结果显示:所有的CGRP免疫阳性神经纤维都是SP免疫阳性,同时所有的SP免疫阳性神经纤维也都是CGRP免疫阳性;CGRP免疫阳性神经纤维的分布和形态与SP免疫阳性神经纤维相一致。结论:P物质和降钙素基因相关肽在大鼠垂体前叶神经纤维内完全共存。     

大鼠心肌氮能神经元及纤维的分布

目的　观测大鼠心脏各部心肌一氧化氮合酶 (nNOS)免疫阳性神经元的形态及分布 ,为探讨氮能神经在心肌的作用提供形态学资料。方法　采用免疫组织化学SABC法显示大鼠心脏nNOS免疫阳性神经元及神经纤维的形态与分布。结果　在大鼠各部心肌内均有氮能神经元和神经纤维分布 ,胞体呈多种形状。nNOS免疫阳性神经纤维呈串珠状、条索状、细线状及不定形状 ,其走行多与肌纤维长轴平行。氮能神经元及纤维的数密度和面密度心房高于心室 ,6月龄高于 1月龄组 (P <0 .0 1)。结论　各部心肌均有氮能神经元及神经纤维 ,心房的含量高于心室。     

P物质、血管活性肠肽和乙酰胆碱能神经在大鼠肠道内的分布及其关系

应用乙酰胆碱酯酶(AChE)组织化学和PAP免疫组织化学方法,比较观察P物质(SP)、血管活性肠肽(VIP)和AChE三种阳性神经元在大鼠十二指肠、空肠、回肠、结肠和直肠内的分布特征及其相互关系。结果显示:SP、VIP、AChE阳性神经神经元和纤维均分布于肠壁各层,从十二指肠、空肠到回肠逐渐增多,但从结肠到直肠则逐渐减少;AChE阳性神经元或纤维在肠壁各层最丰富,其中VIP以粘膜层和粘膜下神经丛较丰富,SP以肠肌丛较丰富;三者的分布密度为AChE>VIP>SP。AChE、SP和VIP阳性神经元胞体及神经纤维在不同肠段的分布密度有明显差异(P<0.05),提示可能与不同肠段肠动力调节功能有关。     

μ阿片受体在大鼠结肠肠肌丛的神经化学特性

目的:探索μ阿片受体(MOR)在大鼠结肠肠肌丛的分布及化学神经解剖学特性。方法:取大鼠结肠右曲和左曲之间肠管行全层铺片,剥离粘膜层、粘膜下层和环形肌层,保留纵行肌层。运用免疫荧光组织化学双重标记技术染色,共聚焦显微镜下观察MOR在大鼠结肠肠肌丛的分布及与一氧化氮合酶(NOS)、肠血管活血肽(VIP)、P物质(SP)或降钙素基因相关蛋白(CGRP)等在肠神经细胞的共存关系。结果:免疫荧光组织化学染色显示:在大鼠结肠肠肌层,MOR阳性细胞聚集形成神经节,其阳性突起穿行于神经节之间形成网格状神经丛。在肠肌丛神经细胞内,可见MOR分别与NOS、VIP、SP、CGRP共存。MOR阳性神经纤维和终末分布于NOS阳性神经细胞周围,也可见部分NOS、VIP阳性神经纤维和终末分布于MOR阳性神经细胞表面。有大量SP和CGRP阳性神经纤维穿行于MOR阳性神经细胞之间并包绕于MOR阳性神经细胞的胞体周围。结论:在结肠的肠肌丛,MOR与抑制性和感觉性神经递质在肠神经细胞有共存,且相互之间有纤维联系,推测MOR可能介导了阿片类调节肠神经细胞内抑制性神经递质的释放,并可能对肠感觉信息的传递有调节作用。     

大鼠心内CGRP免疫话性神经细胞——免疫组织化学PAP法研究

降钙素基因相关肽(CGRP)是由37个氨基酸组成的肽。它广泛存在于中枢和周围神经系统中。目前,国外对心血管系统的CGRP免疫活性神经纤维已有报道,但对心脏CGRP免疫活性神经细胞仅见一篇报道。而国内尚未见到达方面的报道。本实验用石蜡切片,PAP免疫组织化学技术(CGRP抗血清浓度为1:5000)研究大鼠心脏神经细胞的CGRP免疫活性。结果表明:在大鼠心眭神经节中确有CGRP免疫活性的神经细胞存在。这种神经细胞为中等大小、位于主动脉根部附近的心房壁内。 CGRP是一个能以神经递质方式起作用的肽类物质。许多学者认为,CGRP可能是除SP以外感觉神经的第二种标志物。因此,我们推测,大鼠心     

小鼠心脏一氧化氮合酶神经纤维的分布

目的：探讨小鼠心脏一氧化氮合酶aninic oxide synthse神经元结构和神经纤维分布。方法：NADPH-d组织化学技术。结果：小鼠心脏神经元主要分为NOS强阳性、中度和弱阳性反应细胞；心脏各部均接受NOS神经支配，其神经纤维多与肌纤维长轴平行走向。心房最丰富，房室结次之，左、右心室最少。心房及房室结的NOS阳性纤维呈串珠状膨大，心室的常为丝状，膨体极少。结论：小鼠心脏NOS神经元包括强阳性、中度和弱阳性反应细胞，NOS阳性纤维在心脏各部的分布和形态均有差异，NO可能作为神经递质和/或神经调节剂在心血流和冲动传导等的神经调控中起作用。     

神经肽Y、P物质在扬子鳄泄殖腔壁内的分布

目的 :观察含神经肽Y(NPY)、P物质 (SP)神经在扬子鳄泄殖腔壁内的分布情况。方法 :免疫组织化学ABC法及免疫荧光法。结果 :扬子鳄泄殖腔壁内SP免疫反应 (SP IR)阳性神经纤维多见于肌层 ,也见于外膜、呈细线状或点线状 ,肌层内的SP IR神经纤维与平滑肌纤维平行走行或构成网络状 ;NPY免疫反应 (NPY IR)神经纤维呈细线状 ,密度较稀 ,主要见于肌层 ,也近似与平滑肌纤维平行走行 ,免疫荧光法还证实 ,肌层内有散在分布的、多呈椭圆形的NPY IR阳性神经元胞体 ,并见有突起与周围的神经纤维形成联系。结论 :扬子鳄的泄殖腔壁也存在有SP、NPY能神经分布     

正常人肩关节囊内CGRP和SP免疫阳性神经纤维的分布

目的探讨粘连性肩关节囊炎(冻结肩)的发病机制。方法用免疫组织化学方法观察人肩关节囊内CGRP和SP免疫阳性神经纤维存在的情况。结果人肩关节囊的纤维层内有CGRP和SP免疫反应阳性神经纤维存在,免疫反应阳性神经纤维数量较多,独立直行或网状交织。结论CGRP和SP免疫阳性神经纤维可能参与了人肩关节囊的伤害性感觉信息的传递。当人肩关节囊发生炎症时,CGRP和SP免疫阳性神经纤维受到刺激,可能是引起粘连性肩关节囊炎时肩痛的原因之一。     

猪食管神经支配的神经化学研究——一氧化氮类及肽类成分在平滑肌中的分布(英文)

采用免疫荧光组织化学技术及迷走神经切断术,探讨猪食管一氧化氮类及肽类神经支配的神经化学特性。在光学显微镜下可观察到肌间神经丛及粘膜下神经丛中有部分神经元呈nNOS、VIP、GAL、NPY、PACAP、L-ENK、SP、5-HT及CB免疫阳性,但未见CGRP及SOM阳性神经元。nNOS及CB免疫阳性产物主要分布于不同的神经元胞体内。将PGP9.5作为神经元胞体的标记物,并采用免疫荧光免疫组织化学双重染色方法,分别观察了PGP9.5与nNOS、VIP、SP的双标情况。结果如下:(1)nNOS免疫阳性神经元约占PGP9.5标记神经元总数的63%,而VIP免疫阳性神经元约占36%,SP免疫阳性神经元约占28%;(2)神经节内神经元的平均数量呈现吻尾方向的递增趋势,且食管腹段神经丛内神经节数量明显高于食管其他部位;(3)食管肌层内VIP/GAL/NPY免疫阳性纤维分布最广,其中部分阳性纤维同时呈nNOS或PACAP免疫阳性;SP和/或L-ENK免疫阳性纤维在粘膜肌层的分布明显多于平滑肌层。CGRP阳性纤维非常少见,这一点不同于对其他动物的观察结果;(4)经一侧迷走神经切断后,肌间神经丛内PACAP及5-HT免疫阳性纤维明显减少,提示这些纤维可能来源于迷走神经;而平滑肌中VIP/GAL/NPY和/或nNOS免疫阳性纤维数量未发现明显变化,可能为内源性来源。     

Distribution of substance P-immunoreactive and calcitonin gene-related peptide-immunoreactive nerves in normal human lungs.

The distribution of substance P (SP)-immunoreactive (IR) and calcitonin gene-related peptide (CGRP)-IR nerves in normal human lungs was immunohistochemically investigated. SP-IR and CGRP-IR nerves were found in lamina propria and adjacent to blood vessels, and also in some of the ganglia and nerve bundles in the submucosal area. Comparison of serial consecutive sections revealed that SP-IR and CGRP-IR nerves represented similar distribution patterns. CGRP-IR nerves were scarce in bronchial smooth muscles, whereas SP-IR nerves were always present therein. Neither SP-IR nor CGRP-IR nerves were found, however, among submucosal glands or in alveolar septa. These localization patterns were closely related to the functions of SP and CGRP in normal lungs.     

Distribution of capsaicin-sensitive substance P- and calcitonin gene-related peptide-immunoreactive nerves in bovine respiratory tract

The distribution of nerve fibers immunoreactive for substance P (SP) and calcitonin gene-related peptide (CGRP) was examined by means of immunohistochemical methods in the respiratory tract from nose to lung of normal and capsaicin-treated cattle. SP- and CGRP-immunoreactive (IR) nerve fibers with varicosities were detected in all portions. They were more numerous in calves than in cows. They were abundant in the nasal and laryngeal mucosae and tracheal bronchus, and few in number in the lung. SP- and CGRP-IR nerve fibers were mainly seen in the epithelium, in connective tissue beneath the epithelium and around blood vessels, and in the glands throughout the respiratory tract. In contrast, SP- and CGRP-IR nerve fibers were sparse in the smooth muscle layer. Capsaicin treatment of neonates caused a remarkable reduction in the number of SP- and CGRP-IR nerve fibers in the respiratory tract of calves. Double immunofluorescence experiments showed the colocalization of SP and CGRP in most of the nerve fibers. The present findings suggest that SP- and CGRP-IR nerve fibers are involved in the regulation of the bovine respiratory tract, and that capsaicin-sensitive SP- and CGRP-IR nerve fibers are sensory neurons of the bovine respiratory tract.     

Distribution of calcitonin gene-related peptide (CGRP), substance P (SP) and galanin (GAL) immunoreactive nerve fibers in the seminal vesicle and prostate of the male sheep.

Double immunohistochemistry was used to determine the occurrence and distribution pattern of nerve fibres immunoreactive to calcitonin gene-related peptide (CGRP), substance P (SP) and galanin (GAL) in seminal vesicles and prostate of the male sheep. Numerous CGRP- and SP-immunoreactive (IR) nerve fibres were found in the mucosal layer and smooth musculature of the seminal vesicles and prostate. In both glands nerve terminals immunoreactive to CGRP were more numerous than SP-IR ones. The majority of CGRP-IR nerve fibers showed colocalization of this peptide and SP. In both layers of the seminal vesicle and prostate, rare nerve terminals immunoreactive to GAL were also found. Immunoreactivity to SP was also found in all GAL-IR nerve fibers. The presence of numerous CGRP- and SP-IR nerve fibers in the seminal vesicle and prostate of the male sheep suggests that these neuropeptides may be involved in the sensory transmission and/or control of smooth muscle contractility. On the other hand, a relatively low number of GAL-IR nerve fibers of the seminal vesicle and prostate suggest that this peptide may act as an anti-nociceptive agent. It cannot be excluded that, in the seminal vesicle, GAL may also be involved in the control of the smooth muscle fiber activity. The possible role of CGRP, SP and GAL in the regulation of functions of the accessory sexual glands needs to be determined in further physiological studies.     

Calcitonin gene-related peptide containing sensory neurons innervating tooth pulp and buccal mucosa of the rat: an immunohistochemical analysis.

Differential peptidergic innervation of the rat buccal mucosa and incisor tooth pulp was demonstrated by experimental immunohistochemistry. In the buccal mucosa, calcitonin gene-related peptide (CGRP)-like immunoreactive (IR) nerve fibres were observed in both the epithelium and submucosal layer. Most of these fibres were also immunoreactive to substance P (SP). On the other hand, SP-IR fibres were less numerous than CGRP-IR fibres in the incisor tooth pulp. Fast blue (FB) was applied to the buccal mucosa and the incisor pulp chamber. Small (less than 20 microns) FB-labelled CGRP-IR neurons were more numerous in the buccal mucosa group, while medium to large neurons (20-50 microns) predominated in the incisor pulp. Following neonatal capsaicin treatment, CGRP- and SP-containing fibres in the buccal mucosa almost totally disappeared, but many CGRP-IR fibres remained in the incisor tooth pulp. The findings suggest that these two oral tissues are innervated by different subgroups of CGRP-IR neurons in the trigeminal ganglia.     

Distribution of calcitonin gene-related peptide-like immunoreactivity in the guinea pig heart

Summary The distribution of calcitonin gene-related peptide like-immunoreactivity (CGRP-IR) within the heart and adjacent blood vessels of the guinea pig was investigated anmunohistochemically by use of the peroxidase-antiperoxidase (PAP) technique. Numerous paravascular and perivascular immunoreactive nerve fibers were localized around the aorta, coronary arteries and their branches down to the teminal vasculature. Arterioles in the atria showed greatest density of immunoreactive varicosities of all blood vessels. The epicardium, endocardium and the conductive system also contained numerous CGRP-IR nerve fibers. In the muocardium the number of immunoreactive varicosities was variable. Many were present in both atria, moderate amounts were seen in the right ventricle and parts of the intraventricular septum, and only a few occurred in the left ventricle. CGRP-IR was infrequently found within intracardial ganglionic cells but was abundantly distributed in the surrounding nerve fibers.Supported by the German Research Foundation (DFG, SFB 90)     

内脏伤害性传入信息向三叉神经脊束间质核的投射

舌咽、迷走神经向三叉神经脊束间质核的投射纤维除传递非伤害性内脏信息者外是否也含内脏伤害性传入？尚未见报道。本文综合应用：（1）福尔马林刺激舌咽、迷走神经支配的一些内脏诱导FOS蛋白表达；（2）抽除外周神经节观察三叉神经脊束间质核内一些与伤害性信息传递有关的神经递质的变化；（3）运用荧光金逆行追踪与免疫组化相结合的双重反应等方法，对大鼠投射至三叉神经脊束间质核的舌烟、迷走神经初级传入纤维中是否含传递伤害性刺激信息的成分进行了研究。结果证明：(1)2％福尔马林刺激大鼠软腭、咽、食道和胃后，三叉神经脊束间质核内有大量FOS阳性神经元表达，这些阳性神经元与舌咽、迷走神经初级传入分布区基本一致。（2）抽除一侧舌咽、谈走神经报及其神经节后，术侧三叉神经脊束间质核内SP、CGRP等和痛传递有关的神经肽及BSI-B_4阳性终末与对侧比较明显减少。（3）荧光金微量注入三叉神经脊束间质核后，在舌咽、迷走神经节的下节内发现被荧光金逆标的细胞中分别有10％和12％呈SP、CGRP阳性，此两种双重阳性细胞分别占SP、CGRP阳性细胞的33．3%和278％。以上结果提示，三叉神经脊束间质核接受舌咽、迷走神经的传入纤维中有一部分传递内脏伤害性信息。     

A co-localization study on the ovine pancreas innervation

The expression of DbetaH and several neuropeptides was investigated in neuronal elements of the ovine pancreas using double immunocytochemical stainings. Immunoreactivities to DbetaH, NPY, VIP and SP were seen to various extents in nerve terminals associated with the acini, islets, ducts, blood vessels, interlobular connective tissue as well as in the neurons of intrapancreatic ganglia. The expression of CGRP was limited to nerve fibers lying in the connective tissue septa, amongst the acini and in close vicinity to the pancreatic blood vessels. Single GRP-positive nerve endings were located around the acini, ducts and in the interlobular connective tissue. With the exception of the ductal system in a co-localization of NPY with DbetaH was frequently found in all regions of the pancreas. Moderately numerous blood vessel-associated VIP-positive nerve fibers as well as the vast majority of VIP-containing intrapancreatic neurons were found to co-express DbetaH. Single SP-immunoreactive (IR) nerve fibers of the exocrine pancreas and interlobular connective tissue as well as SP-positive intrapancreatic neurons additionally showed the presence of DbetaH. The co-localization of VIP and NPY was found in nerve terminals located around the blood vessels and acini, in the connective tissue septa as well as in numerous pancreatic neuronal perikarya. Rare nerve terminals located between the acini and around small blood vessels as well as several neurons of intrapancreatic ganglia were VIP-IR/ SP-IR. Simultaneous expression of SP and CGRP was found in nerve fibers supplying large pancreatic arteries and veins, interlobular connective tissue and, occasionally, around the acini. Throughout the pancreas the population of CGRP-positive nerve endings showed lack of VIP and NPY. In a moderate number of GRP-containing nerve fibers, a co-expression of NPY was noted. Nerve terminals containing both GRP and VIP were detected sporadically, whereas none of the GRP-positive nerve terminals showed expression of SP. We conclude that the presented noradrenergic as well as peptidergic innervation patterns of the ovine pancreas are species-dependent. On the basis of the occurrence of DbetaH, NPY, VIP and SP (alone or in combination) in pancreatic neuronal elements we can suggest that these substances presumably act as regulators of the endocrine and/or exocrine pancreas. Involvement of CGRP and GRP in the ovine pancreas physiology seems to be of minor importance. The co-localization study indicated that the pancreas of the sheep is innervated from several sources including intrinsic as well as extrinsic ganglia.     

Calcitonin gene-related peptide-immunoreactive nerve fibers in mandibular periosteum of rat: evidence for primary afferent origin

Peptidergic neurons may play a role in the local regulation of bone mineralization. The neuropeptide vasoactive intestinal peptide (VIP) increases bone resorption in vitro, while calcitonin gene-related peptide (CGRP) has been shown to inhibit bone resorption in vitro. We have previously reported that sympathetic nerves with VIP-immunoreactivity innervate bone and periosteum. In the present study we sought to determine if CGRP fibers, like VIP fibers, exist in periosteum and what their origin might be. In whole-mount preparations of mandibular periosteum from rat, CGRP- and VIP-immunoreactive (IR) nerve fibers were present as networks within the periosteum. In preparations using two-color immunofluorescence, most CGRP-IR fibers were also immunoreactive for substance P (SP). In rats in which the subperiosteal space subjacent to the mandibular molars was injected with Fast blue or Fluoro-gold, retrogradely labeled cells were seen in ipsilateral trigeminal ganglia, superior cervical ganglia, and nodose ganglia. Individual cells labeled with both CGRP immunoreactivity and retrograde tracer were seen only in the mandibular portion of the trigeminal ganglion. These data suggest that CGRP-IR nerve fibers in periosteum may be of primary afferent origin. Given the reported effects of CGRP on bone mineralization, the present results suggest that primary afferent nerves containing CGRP and SP, as well as sympathetic nerves containing VIP, may play a role in focal bone remodeling.     

Distribution of calcitonin gene-related peptide immunoreactive nerve fibers in the human submandibular gland

The indirect immunofluorescence technique was used to study the distribution of calcitonin gene-related peptide (CGRP) in human submandibular gland. A relatively low number of thin varicose fibers with intense immunofluorescence for CGRP was seen in samples from seven glands. These CGRP-immunoreactive (CGRP-IR) nerve fibers were mainly seen around or in close contact with intra- and interlobular blood vessels. Some CGRP-IR nerve fibers were also found in association with intra- and interlobular salivary ducts and a few around the submandibular acini. By visual estimation there was no difference in the density of CGRP-IR nerve fibers between specimens of recurrent duct obstruction and laryngeal carcinoma. The present results show that the distribution of CGRP-IR nerve fibers in the stroma and in the glandular secretory elements of the human submandibular gland is quite similar to that seen in the rat and the ferret, which have been reported earlier. Furthermore, the regional distribution of CGRP-IR fibers in the human submandibular gland suggests that CGRP has a physiological role in the regulation of salivary gland function in human salivary glands, e.g. blood flow and secretion.     

Pituitary adenylatecyclase-activating polypeptide-immunoreactive nerve fibers in the rat epiglottis and pharynx

The distribution of pituitary adenylatecyclase-activating polypeptide-immunoreactive (PACAP-IR) nerve fibers was studied in the rat epiglottis and pharynx. PACAP-IR nerve fibers were located beneath the mucous epithelium, and occasionally penetrated the epithelium. These nerve fibers were abundant on the laryngeal side of the epiglottis and in the dorsal and lateral border region between naso-oral and laryngeal parts of the pharynx. PACAP-IR nerve fibers were also detected in taste buds within the epiglottis and pharynx. In addition, many PACAP-IR nerve fibers were found around acinar cells and blood vessels. The double immunofluorescence method demonstrated that distribution of PACAP-IR nerve fibers was similar to that in CGRP-IR nerve fibers in the epithelium and taste bud. However, distributions of PACAP-IR and CGRP-IR nerve fibers innervating mucous glands and blood vessels were different. The retrograde tracing method also demonstrated that PACAP and CGRP were co-expressed by vagal and glossopharyngeal sensory neurons innervating the pharynx. These findings suggest that PACAP-IR nerve fibers in the epithelium and taste bud of the epiglottis and pharynx which originate from the vagal and glossopharyngeal sensory ganglia include nociceptors and chemoreceptors. The origin of PACAP-IR nerve fibers which innervate mucous glands and blood vessels may be the autonomic ganglion.